KCNK5 is functionally down-regulated upon long-term hypotonicity in Ehrlich ascites tumor cells

Signe Skyum Kirkegaard, Tune Wulff, Steen Gammeltoft, Else Kay Hoffmann

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Abstract

Background/Aims: Regulatory volume decrease (RVD) in response to acute cell swelling is well described and KCNK5 (also known as TASK-2 or K 2P 5.1) has been shown to be the volume sensitive K + channel in Ehrlich cells. Very little is, on the other hand, known about the effects of long-term hypotonicity on expression and function of KCNK5, thus we have investigated the effect of long-term hypotonicity (24h - 48h) on KCNK5 in Ehrlich cells on the mRNA, protein and physiological levels. Methods: Physiological effects of long-term hypotonicity were measured using patch-clamp and Coulter counter techniques. Expression patterns of KCNK5 on mRNA and protein levels were established using real-time qPCR and western blotting respectively. Results: The maximum swelling-activated current through KCNK5 was significantly decreased upon 48h of hypotonicity and likewise the RVD response was significantly impaired after both 24 and 48h of hypotonic stimulation. No significant differences in the KCNK5 mRNA expression patterns between control and stimulated cells were observed, but a significant decrease in the KCNK5 protein level 48h after stimulation was found. Conclusion: The data suggest that the strong physiological impairment of KCNK5 in Ehrlich cells after long-term hypotonic stimulation is predominantly due to down-regulation of the KCNK5 protein synthesis.

OriginalsprogEngelsk
TidsskriftCellular Physiology and Biochemistry
Vol/bind32
Udgave nummer5
Sider (fra-til)1238-1246
Antal sider9
ISSN1015-8987
DOI
StatusUdgivet - dec. 2013

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